浙江地区鼬獾和犬源狂犬病病毒分离株全基因组测序与分析  

Sequencing and Analysis of Complete Genome of Rabies Viruses Isolated from Chinese Ferret-Badger and Dog in Zhejiang Province

作　　者：雷永良[1] 王晓光[1] 陶晓燕[2] 李浩[2] 孟胜利[3] 陈秀英[1] 柳付明[1] 叶碧峰[1] 唐青[2] 

机构地区：[1]丽水市疾病预防控制中心,浙江323000 [2]中国疾病预防控制中心病毒病预防控制所,北京100052 [3]武汉生物制品研究所,湖北430060

出　　处：《病毒学报》2010年第1期45-52,共8页Chinese Journal of Virology

基　　金：公益性行业(农业)科研专项(200803014);国家863项目(SQ2006AA02Z112882);2009年浙江省科技计划项目(2009C33166)

摘　　要：测定浙江地区狂犬病病毒分离株（鼬獾和犬）全基因组序列,从分子水平对病毒进行遗传变异特征分析,了解狂犬病病毒在浙江的流行和变异情况以及目前浙江流行株的遗传学背景,以丰富中国狂犬病病毒街毒流行株的全基因组信息。脑内接种1-2日乳鼠分离狂犬病病毒,RT-PCR反应测定浙江地区狂犬病病毒分离株全基因组核苷酸序列,并进行编码蛋白和序列相似性比较及种系发生分析。测序获得狂犬病病毒浙江淳安鼬獾分离株F02、F04和松阳犬分离株D01、D02全基因组核苷酸序列信息：基因组全长11 923和11 925 nts,前导序列Leader长58nts,5个ORF为：NP（1 353 nts）; PP（894 nts）; MP（609 nts）; GP（1 575 nts）; LP（6 386 nts）,N-P-M-G间隔序列长2、5、5 nts; G-L基因间的伪基因Ψ长423 nts; Trailer尾长70 nts。核酸BLAST及多重序列比对分析显示浙江地区4个狂犬病病毒分离株的全基因组序列的组成和结构符合弹状病毒科狂犬病病毒属的特征; 中国毒株之间特别是浙江同种动物狂犬病病毒之间各个基因区域核苷酸与氨基酸序列相似性最高,浙江病毒全基因组序列编码蛋白氨基酸序列相似性高于核苷酸序列相似性,说明蛋白质编码基因的核苷酸变异大多属于同义突变; 浙江病毒负链RNA基因组5个基因编码氨基酸的长度没有变异,5个编码蛋白仅表现较少的序列变化; 浙江病毒与本研究选择的代表性引用街毒株或者来自街毒的减毒株的变异位点和变异类型相似,多重序列相似性的比较和种系发生分析显示所分离的狂犬病病毒浙江街毒株均属于基因1型,具有较独特的中国地域性特点,故本研究中的浙江地区分离株极有可能是自然界中固有的街毒株。Based on sequencing the full-length genomes of four Chinese Ferret-Badger and dog, we analyze the properties of rabies viruses genetic variation in molecular level, get the information about rabies viruses prevalence and variation in Zhejiang, and enrich the genome database of rabies viruses street strains isolated from China. Rabies viruses in suckling mice were isolated, overlapped fragments were amplified by RTPCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses from Chinese Ferret-Badger, dog, sika deer, vole, used vaccine strain were determined. The four full-length genomes were sequenced completely and had the same genetic structure with the length of 11, 923 nts or 11, 925 nts including 58 nts-Leader, 1353 nts-NP, 894 nts-PP, 609 nts-MP, 1575 nts-GP, 6386 nts-LP, and 2, 5, 5 nts-intergenic regions（IGRs）, 423 nts-Pseudogene-like sequence （ψ）, 70 nts-Trailer. The four full-length genomes were in accordance with the properties of Rhabdoviridae Lyssa virus by BLAST and muhi-sequence alignment. The nucleotide and amino acid sequences among Chinese strains had the highest similarity, especially among animals of the same species. Of the four fulllength genomes, the similarity in amino acid level was dramatically higher than that in nucleotide level, so the nucleotide mutations happened in these four genomes were most synonymous mutations. Compared with the reference rabies viruses, the lengths of the five protein coding regions had no change, no recombination, only with a few point mutations. It was evident that the five proteins appeared to be stable. The variation sites and types of the four genomes were similar to the reference vaccine or street strains. And the four strains were genotype 1 according to the multi-sequence and phylogenetic analyses, which possessed the distinct district characteristics of China. Therefore, these four rabies viruses are likely to be street viruses already existing in the natural world.

关 键 词：狂犬病病毒 鼬獾 犬 全基因组 序列分析 

分 类 号：R373.9[医药卫生—病原生物学]